Canine Epithelial Skin Tumours: Expression of the Stem Cell Markers Lgr5, Lgr6 and Sox9 in Light of New Cancer Stem Cell Theories

Evidence is accumulating that tumour development is driven by cancer stem cells (CSCs). In order to understand the presence and potential contribution of stem cells (SCs) as tumour-initiating cells in canine cutaneous tumours, we selected three putative SC markers (Lgr5, Lgr6 and Sox9) and investigated their expression pattern, level of protein and mRNA expression, in 43 canine hair follicle (HF) and 18 canine cutaneous epidermal tumours by immunohistochemistry and qRT-PCR, using normal skin samples as controls. Lgr5 protein expression was not detected in epidermal and HF tumours; however, Lgr5 mRNA overexpression was evident in some HF tumours. Sox9 was expressed in several tumour cases, both at the protein and mRNA level. The Lgr6 antibody tested was not suitable for formalin-fixed paraffin-embedded tissue samples, but Lgr6 gene showed higher expression in several samples of both HF and epidermal tumours compared with normal skin. Significantly higher mRNA expression levels of the three SC markers were found in trichoblastomas (TB) compared with basal cell carcinomas (BCC). The present results indicated that canine HF and epidermal tumours might have common tumour-initiating cells. The mRNA expression of the three selected SC markers, especially Lgr5, could be potentially useful in the distinction between canine TB and BCC.

[1]  C. Blanpain,et al.  Stem cell dynamics, migration and plasticity during wound healing , 2019, Nature Cell Biology.

[2]  S. Skvortsov,et al.  Therapy resistance mediated by cancer stem cells. , 2018, Seminars in cancer biology.

[3]  V. Grieco,et al.  Survivin and Sox9: Potential Stem Cell Markers in Canine Normal, Hyperplastic, and Neoplastic Canine Prostate , 2018, Veterinary pathology.

[4]  J. Nör,et al.  Expression of Cancer Stem Cell Biomarkers in Human Head and Neck Carcinomas: a Systematic Review , 2018, Stem Cell Reviews and Reports.

[5]  H. Nakayama,et al.  Hierarchical Cluster Analysis of Cytokeratins and Stem Cell Expression Profiles of Canine Cutaneous Epithelial Tumors , 2018, Veterinary pathology.

[6]  Sevim Baykal Koca,et al.  CD10, TDAG51, CK20, AR, INSM1, and Nestin Expression in the Differential Diagnosis of Trichoblastoma and Basal Cell Carcinoma , 2018, International journal of surgical pathology.

[7]  C. Ambrósio,et al.  In vitro identification of a stem cell population from canine hair follicle bulge region. , 2018, Tissue & cell.

[8]  P. Harms,et al.  Histologic Mimics of Basal Cell Carcinoma. , 2017, Archives of pathology & laboratory medicine.

[9]  H. Clevers,et al.  Cancer stem cells revisited , 2017, Nature Medicine.

[10]  A. Balmain,et al.  Lgr6 is a stem cell marker in mouse skin squamous carcinomas , 2017, Nature Genetics.

[11]  A. Cuddihy,et al.  Long-Lived Epidermal Cancer-Initiating Cells , 2017, International journal of molecular sciences.

[12]  Z. Modrušan,et al.  Stem cell plasticity enables hair regeneration following Lgr5+ cell loss , 2017, Nature Cell Biology.

[13]  S. Haan,et al.  Tumor-Initiating Cells: a criTICal review of isolation approaches and new challenges in targeting strategies , 2017, Molecular Cancer.

[14]  W. Kim,et al.  Distinct expression profile of stem cell markers, LGR5 and LGR6, in basaloid skin tumors , 2017, Virchows Archiv.

[15]  H. Clevers,et al.  Inactivation of TGFβ receptors in stem cells drives cutaneous squamous cell carcinoma , 2016, Nature Communications.

[16]  M. Welle,et al.  Stem Cell-Associated Marker Expression in Canine Hair Follicles , 2016, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[17]  S. Linnarsson,et al.  Dynamics of Lgr6+ Progenitor Cells in the Hair Follicle, Sebaceous Gland, and Interfollicular Epidermis , 2015, Stem Cell Reports.

[18]  G. Sironi,et al.  Sox9 Expression in Canine Epithelial Skin Tumors , 2015, European journal of histochemistry : EJH.

[19]  L. Bongiovanni,et al.  On the role of survivin as a stem cell biomarker of canine hair follicle and related tumours. , 2014, Veterinary dermatology.

[20]  Xuelin Huang,et al.  An improvement of the 2ˆ(-delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. , 2013, Biostatistics, bioinformatics and biomathematics.

[21]  L. Mechelli,et al.  The contribution of stem cells to epidermal and hair follicle tumours in the dog. , 2013, Veterinary dermatology.

[22]  S. Aerts,et al.  Adult interfollicular tumour-initiating cells are reprogrammed into an embryonic hair follicle progenitor-like fate during basal cell carcinoma initiation , 2012, Nature Cell Biology.

[23]  Tetsuro Kobayashi,et al.  Isolation and characterization of stem cell-enriched human and canine hair follicle keratinocytes. , 2012, Methods in molecular biology.

[24]  S. D’Egidio,et al.  β-catenin in canine skin: immunohistochemical pattern of expression in normal skin and cutaneous epithelial tumours. , 2011, Journal of comparative pathology.

[25]  Tetsuro Kobayashi,et al.  Canine follicle stem cell candidates reside in the bulge and share characteristic features with human bulge cells. , 2010, The Journal of investigative dermatology.

[26]  D. Krahl,et al.  Sox9, more than a marker of the outer root sheath: spatiotemporal expression pattern during human cutaneous embryogenesis , 2010, Journal of cutaneous pathology.

[27]  Tetsuro Kobayashi,et al.  Canine hair-follicle keratinocytes enriched with bulge cells have the highly proliferative characteristic of stem cells. , 2009, Veterinary dermatology.

[28]  L. Bongiovanni,et al.  Apoptosis and anti-apoptotic heat shock proteins in canine cutaneous infundibular keratinizing acanthomas and squamous cell carcinomas. , 2008, Veterinary dermatology.

[29]  E. Fuchs,et al.  Hair follicle stem cells are specified and function in early skin morphogenesis. , 2008, Cell stem cell.

[30]  P. Chambon,et al.  Cutaneous cancer stem cell maintenance is dependent on β-catenin signalling , 2008, Nature.

[31]  V. Vidal,et al.  SOX9 expression is a general marker of basal cell carcinoma and adnexal‐related neoplasms , 2008, Journal of cutaneous pathology.

[32]  F. Mercati,et al.  CD34 glycoprotein identifies putative stem cells located in the isthmic region of canine hair follicles. , 2006, Veterinary dermatology.

[33]  J. Ortonne,et al.  Sox9 Is Essential for Outer Root Sheath Differentiation and the Formation of the Hair Stem Cell Compartment , 2005, Current Biology.

[34]  I. Kurokawa,et al.  Solid basal cell epithelioma (BCE) possibly originates from the outer root sheath of the hair follicle. , 1993, Acta dermato-venereologica.

[35]  L. Bongiovanni,et al.  SOX9 and CK15 stem cell markers in canine sebaceous lesions , 2015 .

[36]  M. Goldschmidt.  Histological classification of epithelial and melanocytic tumors of the skin of domestic animals , 1998 .